title: Cloning, expression, purification and characterization of an iron-dependent regulator protein from Thermobifida fusca.
abstract: Iron-dependent regulators (IdeRs) control the transcription of a variety of genes associated with iron homeostasis in Gram-positive bacteria. In this study we report the cloning of a putative IdeR gene from the moderate thermophile Thermobifida fusca into the pET-21a(+) expression vector. The expressed protein, Tf-IdeR, was purified using immobilized metal affinity and size-exclusion chromatography, and yielded approximately 12-16 mg of protein per liter of culture. The purified Tf-IdeR protein binds the tox operator sequence in the presence of divalent metal ions. Two Tf-IdeR binding sites were identified in the T. fusca genome upstream of a putative enterobactin exporter and a putative ABC-type multidrug transporter.
authors.affiliation: Department of Chemistry, Wabash College, 301 W. Wabash Ave., Crawfordsville, IN 47933, USA. Electronic address: jbgrange13@wabash.edu.
authors.forename: Joseph B
authors.initials: JB
authors.lastname: Granger
authors.forename: Zeyu
authors.initials: Z
authors.lastname: Lu
authors.forename: Jordan B
authors.initials: JB
authors.lastname: Ferguson
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Santa Maria
authors.forename: Walter R P
authors.initials: WR
authors.lastname: Novak
mesh_terms: D000192:Actinomycetales / Q000235:genetics*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016415:Sequence Alignment
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D012097:Repressor Proteins
keywords: DNA-binding; DtxR; IdeR; Iron-dependent regulator protein; Protein purification; Thermobifida fusca